Long-Term Fluorescence Recovery After Photobleaching (FRAP)

Abstract

Numerous models have been developed for the analysis of fluorescence recovery after photobleaching (FRAP), by which intracellular diffusion and turnover rate are quantitatively evaluated. FRAP analyses typically focus on such events that occur within several minutes, but to precisely evaluate a slow turnover rate of particularly actin stress fibers, achieving long-term FRAP observations of more than 10 min is necessary. In such long-term observations, the effect of intracellular advection is no longer ignored, which motivated us to develop a novel method to decouple the multiple factors associated with the long FRAP response. This method allows us to distinguish the origin of mechanobiological responses of stress fibers that come from either the level of individual actin filaments or that of actin monomers.

Keywords: Advection; Chemical reaction; Confocal laser scanning microscopy; Cytoskeleton; Fluorescence recovery after photobleaching; Live-cell imaging; Stress fiber; Turnover.